生物技术通报2017，Vol.33Issue(11)：166-173,8.DOI:10.13560/j.cnki.biotech.bull.1985.2017-0451
大黄鱼Lc-UBE2D4基因的克隆及其表达分析
Cloning and Expression Profiles of Lc-UBE2D4 Gene from Large Yellow Croaker Larimichthys crocea
摘要
Abstract
Protein uhiquitination is a fundamental post-translational modification event that serves as a signaling function in diverse biological processes,including cell-cycle progression,cell apoptosis,gene transcriptional regulation and expression,etc.In these processes of ubiquitin-proteasome pathway,ubiquitin-conjugating enzyme E2 is of pivotal importance that mediates more than 80 percent of protein degradation of eukaryotic cells,and it plays a key role on the specific selection of degraded of target protein.In this study,a homologous gene fragment of ubiquitin-conjugating enzymes E2 D4 (Lc-UBE2D4) was screened from previous constructed normalized gonad cDNA library of large yellow croaker Larimichthys crocea was obtained,and its full length cDNA was cloned by using SMART-RACE technology,and then the differential expression profiles in different tissues and different gonadal development stages were analyzed using quantitative real time PCR (qRT-PCR).The full length cDNA of Lc-UBE2D4 gene was of 798 bp,with 441 bp of ORF encoding a protein of 147 amino acids,and it contained the highly conserved ubiquitin-conjugating enzymes UBC domain and an active site cysteine.The qRT-PCR results revealed that the highest expression level of Lc-UBE2D4 presented in spleen and testis which were extremely higher than those in other tissues (P ＜ 0.01),and its expression level in ovary was significantly higher than other organs (excluding spleen and testis) (P ＜ 0.05).Moreover,its expression levels in three stages of testis were higher than in the same growth stages of ovary (P ＜ 0.05).These results suggested that Lc-UBE2D4 played essential roles in the gonad development,especially in testis development.Meanwhile,Lc-UBE2D4 may be involved in immune function due to highest expression in spleen.关键词
泛素化/Lc-UBE2D4/大黄鱼/组织表达/性腺发育Key words
ubiquitination/Lc-UBE2D4/large yellow croaker/tissue expression/gonad development引用本文复制引用
韩坤煌,周鹏,邹志华,张子平,王艺磊..大黄鱼Lc-UBE2D4基因的克隆及其表达分析[J].生物技术通报,2017,33(11):166-173,8.基金项目
福建省自然科学基金项目(2015J06019),福建省STS项目(2016T3041),福建省海洋生物资源开发利用协同创新中心产学研基金(FJMBIO1502),大黄鱼育种国家重点实验室开放课题基金(LYC2016RS02),厦门南方海洋研究中心资助项目(14GZP75NF39) （2015J06019）
